Behavioral and molecular response of the insect parasitic nematode Steinernema carpocapsae to plant volatiles.

Entomopathogenic nematodes (EPNs) use the chemical cues emitted by insects and insect-damaged plants to locate their hosts. Steinernema carpocapsae, a species of EPN, is an established biocontrol agent used against insect pests. Despite its promising potential, the molecular mechanisms underlying its ability to detect plant volatiles remain poorly understood. In this study, we investigated the response of S. carpocapsae infective juveniles (IJs) to 8 different plant volatiles. Among these, carvone was found to be the most attractive volatile compound. To understand the molecular basis of the response of IJs to carvone, we used RNA-Seq technology to identify gene expression changes in response to carvone treatment. Transcriptome analysis revealed 721 differentially expressed genes (DEGs) between carvone-treated and control groups, with 403 genes being significantly upregulated and 318 genes downregulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the responsive DEGs to carvone attraction were mainly involved in locomotion, localization, behavior, response to stimulus, and olfactory transduction. We also identified four upregulated genes of chemoreceptor and response to stimulus that were involved in the response of IJs to carvone attraction. Our results provide insights into the potential transcriptional mechanisms underlying the response of S. carpocapsae to carvone, which can be utilized to develop environmentally friendly strategies for attracting EPNs.

The Effect of Nitrogen Annealing on the Resistive Switching Characteristics of the W/TiO2/FTO Memory Device.

In this paper, the effect of nitrogen annealing on the resistive switching characteristics of the rutile TiO2 nanowire-based W/TiO2/FTO memory device is analyzed. The W/TiO2/FTO memory device exhibits a nonvolatile bipolar resistive switching behavior with a high resistance ratio (RHRS/RLRS) of about two orders of magnitude. The conduction behaviors of the W/TiO2/FTO memory device are attributed to the Ohmic conduction mechanism and the Schottky emission in the low resistance state and the high resistance state, respectively. Furthermore, the RHRS/RLRS of the W/TiO2/FTO memory device is obviously increased from about two orders of magnitude to three orders of magnitude after the rapid nitrogen annealing treatment. In addition, the change in the W/TiO2 Schottky barrier depletion layer thickness and barrier height modified by the oxygen vacancies at the W/TiO2 interface is suggested to be responsible for the resistive switching characteristics of the W/TiO2/FTO memory device. This work demonstrates the potential applications of the rutile TiO2 nanowire-based W/TiO2/FTO memory device for high-density data storage in nonvolatile memory devices.

Investigating the Regulatory Mechanism of the Sesquiterpenol Nerolidol from a Plant on Juvenile Hormone-Related Genes in the Insect Spodoptera exigua.

Various plant species contain terpene secondary metabolites, which disrupt insect growth and development by affecting the activity of juvenile hormone-degrading enzymes, and the juvenile hormone (JH) titers maintained in insects. Nerolidol, a natural sesquiterpenol belonging to the terpenoid group, exhibits structural similarities to insect JHs. However, the impact of nerolidol on insect growth and development, as well as its underlying molecular mechanism, remains unclear. Here, the effects of nerolidol on Spodoptera exigua were investigated under treatment at various sub-lethal doses (4.0 mg/mL, 1.0 mg/mL, 0.25 mg/mL). We found that a higher dose (4.0 mg/mL) of nerolidol significantly impaired the normal growth, development, and population reproduction of S. exigua, although a relatively lower dose (0.25 mg/mL) of nerolidol had no significant effect on this growth and development. Combined transcriptome sequencing and gene family analysis further revealed that four juvenile hormone esterase (JHE)-family genes that are involved in juvenile hormone degradation were significantly altered in S. exigua larvae after nerolidol treatment (4.0 mg/mL). Interestingly, the juvenile hormone esterase-like (JHEL) gene Sexi006721, a critical element responsive to nerolidol stress, was closely linked with the significant augmentation of JHE activity and JH titer in S. exigua (R2 = 0.94, p < 0.01). Taken together, we speculate that nerolidol can function as an analog of JH by modulating the expression of the enzyme genes responsible for degrading JH, resulting in JH disorders and ultimately disrupting the development of insect larvae. This study ultimately provides a theoretical basis for the sustainable control of S. exigua in the field whilst proposing a new perspective for the development of novel biological pesticides.

In Situ Low-Temperature Carbonization Capping of LiFePO4 with Coke for Enhanced Lithium Battery Performance.

Lithium batteries incorporating LiFePO4 (LFP) as the cathode material have gained significant attention in recent research. However, the limited electronic and ionic conductivity of LFP poses challenges to its cycling performance and overall efficiency. In this study, we address these issues by synthesizing a series of LiFePO4/carbon (LFP/C) composites through low-temperature carbonization coating of LFP in the presence of Coke as the carbon source. The resulting lithium batteries utilizing LFP/C as the cathode material exhibited impressive discharge specific capacities of 148.35 mA·h/g and 126.74 mA·h/g at 0.1 C and 1 C rates, respectively. Even after 200 cycles of charging and discharging, the capacities remained remarkably high, with values of 93.74% and 97.05% retention, showcasing excellent cycling stability. Notably, the LFP/C composite displayed exceptional rate capability, and capacity retention of 99.27% after cycling at different multiplication rates. These findings underscore the efficacy of in situ low-temperature carbonization capping of LFP with Coke in significantly improving both the cycling stability and rate capability of lithium batteries.

Tunable Resistive Switching Behaviors and Mechanism of the W/ZnO/ITO Memory Cell.

A facile sol-gel spin coating method has been proposed for the synthesis of spin-coated ZnO nanofilms on ITO substrates. The as-prepared ZnO-nanofilm-based W/ZnO/ITO memory cell showed forming-free and tunable nonvolatile multilevel resistive switching behaviors with a high resistance ratio of about two orders of magnitude, which can be maintained for over 103 s and without evident deterioration. The tunable nonvolatile multilevel resistive switching phenomena were achieved by modulating the different set voltages of the W/ZnO/ITO memory cell. In addition, the tunable nonvolatile resistive switching behaviors of the ZnO-nanofilm-based W/ZnO/ITO memory cell can be interpreted by the partial formation and rupture of conductive nanofilaments modified by the oxygen vacancies. This work demonstrates that the ZnO-nanofilm-based W/ZnO/ITO memory cell may be a potential candidate for future high-density, nonvolatile, memory applications.

A Facile Hydrothermal Synthesis and Resistive Switching Behavior of α-Fe2O3 Nanowire Arrays.

A facile hydrothermal process has been developed to synthesize the α-Fe2O3 nanowire arrays with a preferential growth orientation along the [110] direction. The W/α-Fe2O3/FTO memory device with the nonvolatile resistive switching behavior has been achieved. The resistance ratio (RHRS/RLRS) of the W/α-Fe2O3/FTO memory device exceeds two orders of magnitude, which can be preserved for more than 103s without obvious decline. Furthermore, the carrier transport properties of the W/α-Fe2O3/FTO memory device are dominated by the Ohmic conduction mechanism in the low resistance state and trap-controlled space-charge-limited current conduction mechanism in the high resistance state, respectively. The partial formation and rupture of conducting nanofilaments modified by the intrinsic oxygen vacancies have been suggested to be responsible for the nonvolatile resistive switching behavior of the W/α-Fe2O3/FTO memory device. This work suggests that the as-prepared α-Fe2O3 nanowire-based W/α-Fe2O3/FTO memory device may be a potential candidate for applications in the next-generation nonvolatile memory devices.

Monodisperse MoS2/Graphite Composite Anode Materials for Advanced Lithium Ion Batteries.

Traditional graphite anode material typically shows a low theoretical capacity and easy lithium decomposition. Molybdenum disulfide is one of the promising anode materials for advanced lithium-ion batteries, which possess low cost, unique two-dimensional layered structure, and high theoretical capacity. However, the low reversible capacity and the cycling-capacity retention rate induced by its poor conductivity and volume expansion during cycling blocks further application. In this paper, a collaborative control strategy of monodisperse MoS2/graphite composites was utilized and studied in detail. MoS2/graphite nanocomposites with different ratios (MoS2:graphite = 20%:80%, 40%:60%, 60%:40%, and 80%:20%) were prepared by mechanical ball-milling and low-temperature annealing. The graphite sheets were uniformly dispersed between the MoS2 sheets by the ball-milling process, which effectively reduced the agglomeration of MoS2 and simultaneously improved the electrical conductivity of the composite. It was found that the capacity of MoS2/graphite composites kept increasing along with the increasing percentage of MoS2 and possessed the highest initial discharge capacity (832.70 mAh/g) when MoS2:graphite = 80%:20%. This facile strategy is easy to implement, is low-cost, and is cosmically produced, which is suitable for the development and manufacture of advance lithium-ion batteries.

Synergistic effect of carboxyl and sulfate groups for effective removal of radioactive strontium ion in a Zr-metal-organic framework.

Rapid removal of radioactive strontium from nuclear wastewater is of great significance for environmental safety and human health. This work reports the effective adsorption of strontium ion in a stable dual-group metal-organic framework, Zr6(OH)14(BDC-(COOH)2)4(SO4)0.75 (Zr-BDC-COOH-SO4), which contains strontium-chelating groups (-COOH and SO4) and a strongly ionizable group (-COOH). Zr-BDC-COOH-SO4 exhibits very rapid adsorption kinetics (<5 min) and a maximum adsorption capacity of 67.5 mg g-1. The adsorption behaviors can be well fitted to the pseudo-second-order model and the Langmuir isotherm model. Further investigations indicate that the adsorption of Sr2+ onto Zr-BDC-COOH-SO4 would not be obviously affected by solution pH and adsorption temperature. The feasible regeneration of the adsorbent was also demonstrated using a simple elution method. Mechanism investigation suggests that free -COOH contributes to the rapid adsorption based on electrostatic interaction, while the introduction of -SO4 significantly enhanced the adsorption capacity. Thus, these results suggest that Zr-BDC-COOH-SO4 is a potential candidate for Sr2+ removal. They also introduce dual groups as an effective strategy for designing high-efficiency adsorbents.

Hydroxyapatite (HA) Modified Nanocoating Enhancement on AZ31 Mg Alloy by Combined Surface Mechanical Attrition Treatment and Electrochemical Deposition Approach.

Hydroxyapatite (HA) nanocoating was electrodeposited on the surface mechanical attrition treated (SMATed) AZ31 magnesium alloy. Phases, morphologies and the adhesion of coating were characterized by X-ray diffraction, scanning electron microscopy (SEM) and 3D optical profiler. The corrosion resistance of the HA coating was tested by potentiodynamic polarization and electrochemical impedance spectroscopy (EIS). The results showed that the HA coating on SMATed sample had a better crystallization than that on original one. The thickness of HA coating increased from 25 to 40 µm. The bonding strength between HA coating and SMATed substrate was higher than that between the coating and untreated counterpart. Potentiodynamic polarization and EIS demonstrated that the corrosion current density of HA coating on SMATed substrate decreased by 30.84% than that on original. The corrosion potential shifted 80.3 mV to the positive direction. The corrosion resistance of coatings on SMATed sample was significantly enhanced. The immersion experiments showed that the HA coatings on SMATed sample exhibited a better biological activity.

Diversity analysis of intestinal microflora between healthy and diarrheal neonatal piglets from the same litter in different regions.

Though the problem of neonatal piglet diarrhea is well known, the differences in the bacterial diversity and community composition between healthy and diarrheal piglets are still unknown. We investigated these differences in neonatal piglets from Jiangxi Province, China. Healthy (H, n = 20) and diarrheal (D, n = 20) piglets were selected from six regions. The fecal microbial communities were analyzed by sequencing the V3V4 region of 16S rRNA gene. We found the ratio of major phyla (Fusobacteria, Bacteroidetes, Firmicutes and Proteobacteria) was >99% of 7 phyla. The overall alpha diversity indices, such as chao, sobs, coverage and Shannon, were not significantly different. Moreover, the relative abundance of the predicted functions was highly similar in the two groups. Our results indicated that Clostridium was divided into two major groups: Clostridium sensu stricto_1 and stricto_2. Sensu stricto_2 was highly abundant in the D group and low abundance in the H group, whereas the results of sensu stricto_1 were opposite. Comparative analyses within the H or D groups showed that Escherichia-Shigella and Streptococcus at the genus level and unclassified Lactobacillus at the species level were significant difference. Comparative analyses of the two groups showed that unclassified Prevotellaceae at the genus level and Fusobacterium mortifierum were significantly different and had high linear discriminant analysis (LDA) scores. The significantly different microbes composition results also existed in the same litter, based on excluding regions influence. These results suggested that piglet diarrhea was closely associated with these microbes. This study provides insights into gut microbial interactions and prevention of piglet diarrhea.

Fluorescence spectra, fluorescence quantum yield and dissociation constant of sarafloxacin.

In this study, the fluorescence spectra of sarafloxacin (SAR) under different pH conditions were investigated to determine the structural changes due to protonation that result from change in pH. At pH < 1.02, SAR exists in the H3 L2+ form for which the maximum fluorescence emission wavelength was about 455 nm. At pH 1.87-4.94, SAR exists in the H2 L+ form in which H3 L2+ loses one proton in the nitrogen molecule at the 1-position in the quinoline ring. Fluorescence intensity was strong and steady and the maximum emission wavelength was 458 nm. At pH 7.14-9.30, the maximum emission wavelengths were gradually blue shifted to 430 nm with increase in pH, here SAR exists in the form of a bipolar ion HL in which H2 L+ loses a carboxyl group proton. At pH > 11.6, HL transforms into anionic L- in which HL loses one proton from the piperazine ring, leading to a decrease in fluorescence intensity, and the maximum emission wavelength was red shifted to approximately 466 nm. The two-step dissociation constant pKa for SAR was calculated, pKa1 was 6.06 ± 0.37 and pKa2 for SAR was 10.53 ± 0.19. In a pH 3.62 buffer solution with quinine sulfate as the reference, the fluorescence quantum yield of SAR at the maximum excitation wavelength of 276 nm was 0.09.

Investigation on the interaction of cefpirome sulfate with lysozyme by fluorescence quenching spectroscopy and synchronous fluorescence spectroscopy.

The reaction mechanism of cefpirome sulfate with lysozyme at different temperatures (298, 310 and 318 K) was investigated using fluorescence quenching and synchronous fluorescence spectroscopy under simulated physiological conditions. The results clearly demonstrated that cefpirome sulfate caused strong quenching of the fluorescence of lysozyme by a static quenching mechanism. The binding constants obtained using the above methods were of the same order of magnitude and very similar. Static electric forces played a key role in the interaction between cefpirome sulfate and lysozyme, and the number of binding sites in the interaction was close to 1. The values of Hill's coefficients were > 1, indicating that drugs or proteins showed a very weakly positive cooperativity in the system. In addition, the conclusions obtained from the two methods using the same equation were consistent. The results indicated that synchronous fluorescence spectrometry could be used to study the binding mechanism between drug and protein, and was a useful supplement to the fluorescence quenching method. In addition, the effect of cefpirome sulfate on the secondary structure of lysozyme was analyzed using circular dichroism spectroscopy.

Investigation on the effect of fluorescence quenching of bovine serum albumin by cefoxitin sodium using fluorescence spectroscopy and synchronous fluorescence spectroscopy.

The reaction mechanism of cefoxitin sodium with bovine serum albumin was investigated using fluorescence spectroscopy and synchronous fluorescence spectroscopy at different temperatures. The results showed that the change of binding constant of the synchronous fluorescence method with increasing temperature could be used to estimate the types of quenching mechanisms of drugs with protein and was consistent with one of fluorescence quenching method. In addition, the number of binding sites, type of interaction force, cooperativity between drug and protein and energy-transfer parameters of cefoxitin sodium and bovine serum albumin obtained from two methods using the same equation were consistent. Electrostatic force played a major role in the conjugation reaction between bovine serum albumin and cefoxitin sodium, and the type of quenching was static quenching. The primary binding site for cefoxitin sodium was sub-hydrophobic domain IIA, and the number of binding sites was 1. The value of Hill's coefficients (nH ) was approximately equal to 1, which suggested no cooperativity in the bovine serum albumin-cefoxitin sodium system. The donor-to-acceptor distance r < 7 nm indicated that static fluorescence quenching of bovine serum albumin by cefoxitin sodium was also a non-radiation energy-transfer process. The results indicated that synchronous fluorescence spectrometry could be used to study the reaction mechanism between drug and protein, and was a useful supplement to the conventional method. Copyright © 2015 John Wiley & Sons, Ltd.

Using resonance light scattering and UV/vis absorption spectroscopy to study the interaction between gliclazide and bovine serum albumin.

At different temperatures (298, 310 and 318 K), the interaction between gliclazide and bovine serum albumin (BSA) was investigated using fluorescence quenching spectroscopy, resonance light scattering spectroscopy and UV/vis absorption spectroscopy. The first method studied changes in the fluorescence of BSA on addition of gliclazide, and the latter two methods studied the spectral change in gliclazide while BSA was being added. The results indicated that the quenching mechanism between BSA and gliclazide was static. The binding constant (Ka ), number of binding sites (n), thermodynamic parameters, binding forces and Hill's coefficient were calculated at three temperatures. Values for the binding constant obtained using resonance light scattering and UV/vis absorption spectroscopy were much greater than those obtained from fluorescence quenching spectroscopy, indicating that methods monitoring gliclazide were more accurate and reasonable. In addition, the results suggest that other residues are involved in the reaction and the mode 'point to surface' existed in the interaction between BSA and gliclazide. Copyright © 2015 John Wiley & Sons, Ltd.

Characteristic Steroids from the South China Sea Gorgonian Muricella sibogae and Their Cytotoxicities.

Twenty-four steroid-based natural products, 9,10-secosteroids (1-10), 1,4-dien-3-one steroids (11-19), and 4-en-3-one steroids (20-24), containing varying side-chains, were isolated from the South China Sea gorgonian Muricella sibogae. The structures of one new 9,10-secosteroid, sibogol D (1), and two new 1,4-dien-3-one steroids, sibogols E and F (11 and 12), were elucidated by extensive spectroscopic analyses and by comparison with the literature data. Cytotoxicities for all the isolates were evaluated against four selected tumor cell lines, HL-60, HCT116, K562, and P388. Compounds 3, 9, and 13 exhibited potent cytotoxic activities against the HL-60 cell line, with IC50 values ranging from 1.27 to 10.05 µM, and compound 3 was also cytotoxic against HCT116 with an IC50 value of 5.8 µM. The bioassay results also indicated a potential relationship between structural patterns and activity. The newly presented series of 1,4-dien-3-one and 4-en-3-one types of steroids relating to the unique 9,10-secosteroids in biogenesis were found in this species for the first time, which is of considerable chemotaxonomic significance.

[The Investigation on the Interaction of Colistin Sulfate with Bovine Serum Albumin with Resonance Light Scattering Spectroscopy].

The interaction between colistin sulfate (CS) with bovine serum albumin in physiological buffer (pH 7.4) was investigated with resonance light scattering spectroscopy at 298, 310, and 318 K. The analysis of data indicated that quenching mechanism of BSA-CS was probably static. The value of n was approximately 1, indicating there was only a single class of binding sites on BSA for CS compounds. The thermodynamic parameters were calculated at different temperatures, implying that the interaction was spontaneous and electrostatic force played major role in the binding between CS and BSA. The values of nH were equal to 1 at different temperatures, indicating there was non-cooperative reaction between BSA and CS. The feasibility of resonance light scattering spectroscopy was verified by fluorescence quenching spectroscopy. The quenching reactive parameters (KSV，Kq，n，Ka) from two methods were similar, suggesting resonance light scattering spectroscopy could be used to study the binding interaction between protein and drugs. Resonance light scattering spectroscopy can be used to explore the substance without intrinsic fluorescence, suggesting that the application of resonance light scattering spectroscopy broadens the understanding of the interaction between small molecules and protein.

Comparative studies on the interaction of cefixime with bovine serum albumin by fluorescence quenching spectroscopy and synchronous fluorescence spectroscopy.

Under simulated physiological conditions, the reaction mechanism between cefixime and bovine serum albumin at different temperatures (293, 303 and 310 K) was investigated using a fluorescence quenching method and synchronous fluorescence method, respectively. The results indicated that the fluorescence intensity and synchronous fluorescence intensity of bovine serum albumin decreased regularly on the addition of cefixime. In addition, the quenching mechanism, binding constants, number of binding sites, type of interaction force and energy-transfer parameters of cefixime with bovine serum albumin obtained from two methods using the same equation were consistent. The results indicated that the synchronous fluorescence spectrometry could be used to study the binding mechanism between drug and protein, and was a useful supplement to the conventional method.

Cloning of Two Acetylcholinesterase Genes and Analysis of Point Mutations Putatively Associated with Triazophos Resistance in Chilo auricilius (Lepidoptera: Pyralidae).

Acetylcholinesterase (AChE) is the target of organophosphate (OP) and carbamate insecticides. Mutations in the AChE gene (ace) leading to decreased insecticide susceptibility is the main resistance mechanism in insects. In this study, two Chilo auricilius acetylcholinesterase genes, designated as Caace1 and Caace2, were cloned using RT-PCR and RACE. Caace1 cDNA is 2534 bp, with ORF of 2082 bp, and it encodes an acetylcholinesterase 1 (CaAChE1) protein comprising a calculated 693 amino acid (aa) residues. Caace2 cDNA contains 2280 bp, with a full-length ORF of 1917 bp, encoding acetylcholinesterase 2 (CaAChE2) comprising a calculated 638 aa residues. At the aa level, CaAChE1 displays the highest similarity (97%) with the Chilo suppressalis AChE1, and CaAChE2 shows the highest similarity with the C. suppressalis AChE2 (99%). From the restriction fragment length polymorphism (RFLP) PCR (RFLP-PCR) analysis, one mutation in Caace1, similar to the ace1 mutation associated with triazophos resistance in C. suppressalis, was detected. Detailed examination of field populations of C. auricilius indicated this resistance mutation in C. auricilius is still quite infrequent. Based on the assay of AChE activity and RFLP-PCR testing, an individual that contains resistance mutation has lower AChE activities, while the individual that does not contain the resistance mutation has higher AChE activities. This study provides a basis for future investigations into the mechanism of OP resistance in C. auricilius, as well as a guidance for C. auricilius control with reasonable choice of pesticides.

Molecular characterization of the piggyBac-like element, a candidate marker for phylogenetic research of Chilo suppressalis (Walker) in China.

BACKGROUND: Transposable elements (TEs, transposons) are mobile genetic DNA sequences. TEs can insert copies of themselves into new genomic locations and they have the capacity to multiply. Therefore, TEs have been crucial in the shaping of hosts' current genomes. TEs can be utilized as genetic markers to study population genetic diversity. The rice stem borer Chilo suppressalis Walker is one of the most important insect pests of many subtropical and tropical paddy fields. This insect occurs in all the rice-growing areas in China. This research was carried out in order to find diversity between C. suppressalis field populations and detect the original settlement of C. suppressalis populations based on the piggyBac-like element (PLE). We also aim to provide insights into the evolution of PLEs in C. suppressalis and the phylogeography of C. suppressalis. RESULTS: Here we identify a new piggyBac-like element (PLE) in the rice stem borer Chilo suppressalis Walker, which is called CsuPLE1.1 (GenBank accession no. JX294476). CsuPLE1.1 is transcriptionally active. Additionally, the CsuPLE1.1 sequence varied slightly between field populations, with polymorphic indels (insertion/deletion) and hyper-variable regions including the identification of the 3' region outside the open reading frame (ORF). CsuPLE1.1 insertion frequency varied between field populations. Sequences variation was found between CsuPLE1 copies and varied within and among field populations. Twenty-one different insertion sites for CsuPLE1 copies were identified with at least two insertion loci found in all populations. CONCLUSIONS: Our results indicate that the initial invasion of CsuPLE1 into C. suppressalis occurred before C. suppressalis populations spread throughout China, and suggest that C. suppressalis populations have a common ancestor in China. Additionally, the lower reaches of the Yangtze River are probably the original settlement of C. suppressalis in China. Finally, the CsuPLE1 insertion site appears to be a candidate marker for phylogenetic research of C. suppressalis.

Studies on the interaction of palmatine hydrochloride with bovine hemoglobin.

The interaction between bovine hemoglobin (BHb) and palmatine hydrochloride (PMT) was investigated at different temperatures using multispectroscopy, as well as the effect of common metal ions (Ca(2+) , Mg(2+) , Zn(2+) , Cu(2+) , Fe(2+) , Fe(3+) , Co(2+) , Ni(2+) ) on the BHb-PMT system. Results showed that the quenching mechanism of PMT on BHb was a static process. The electrostatic force played an important role in the conjugation reaction between BHb and PMT. The order of magnitude of the binding constants (Ka ) was 10(4) , and the number of binding sites (n) in the binary system was ~ 1. The binding distance (r) was ~ 2.44 nm and the primary binding for PMT was located at ß-37 tryptophan in the hydrophobic cavity of BHb. In addition, the Hill's coefficients were ~ 1. Synchronous and circular dichroism spectra revealed that the microenvironment and the conformation of BHb were changed during the binding reaction.